File S1 - Ubiquitin Ligase gp78 Targets Unglycosylated Prion Protein PrP for Ubiquitylation and Degradation

Supporting Figures. Figure S1. ugPrP binds gp78 but not HsHrd1. (A) gp78 immunoprecipitates ugPrP. Plsamids expressing myc-tagged gp78 and/or ugPrP as indicated were transfected into HEK293 cells. Proteins were extracted and immunoprecipitated with beads coated with myc antibody. Immunoprecipitates were separated on SDS-PAGE, and probed with anti- 3F4 (top panel) or myc antibody (middle panel). The amounts of ugPrP in cell extracts were evaluated and presented in lower panel. (B) ugPrP does not immunoprecipitate HsHrd1. Proteins were extracted from cells expressing myc-tagged HsHrd1 and ugPrP. The indicated immunoprecipitations and immunoblottings were carried out as described above in (A). Figure S2. Quantitation of the data in Figure 3B. The experiments were done at least three times, and the average values with standard deviation are shown. Figure S3. Quantitation of the data shown in Figure 4B. The stability measurements were done more than three times, and the average values with standard deviation are shown. (PDF)